Beurteilung der Lungengesundheit von Schweinen aus verschiedenen ökologischen Haltungssystemen mittels BAL und Lungenhistologie

Im vorliegenden Forschungsvorhaben wurde die Lungengesundheit von Schweinen in Abhängigkeit von verschiedenen Haltungs- und Managementbedingungen beschrieben. Die Untersuchung erfolgte an Mastschweinen verschiedener Herkünfte (konventionell – ökologischer Tiefstreustall – Stall nach EU-Öko-VO – Freilandhaltung) und an Tieren unterschiedlichen Alters. In der konventionellen Schweinemast handelte es sich hierbei um Ferkel, Läufer und Mastschweine derselben Herkunft, in der ökologischen Schweinemast um Läufer, Mastschweine und XXL-Schweine von jeweils zwei Beständen. Letztere Untersuchung diente vor allem der Abbildung möglicher Effekte von Unterschieden im betrieblichen und Hygienemanagement auf die Tiergesundheit in den beiden ökologischen Beständen. Der Gesundheitszustand der Tiere mit Betonung des Lungenbefundes wurde über Schlachtkörperuntersuchungen, bronchoalveoläre Lavage (BAL) und histologische Untersuchung makroskopisch auffälliger Lungenbefunde erfasst. Allgemein wurde festgestellt, dass die Tiere im Zeitverlauf und in Abhängigkeit von den Haltungsbedingungen Zeichen einer zunehmenden entzündlichen Veränderung im Bereich der Atemwege zeigten (BAL: prozentuale Reduktion der Makrophagen bei gleichzeitigem relativen Anstieg der Lymphozyten). Die histologische Untersuchung wies auf ein unterschiedlich ausgeprägtes v.a. viral bedingtes infektiöses Geschehen hin, das möglicherweise durch eine in unterschiedlichem Ausmaß bestehende irritative Schädigung der Atemwege gefördert wurde. Sowohl bei Untersuchung der Mastschweine als auch bei Probenahme bei unterschiedlich alten Tieren wurde deutlich, dass den Managementfaktoren (Entwurmung, Impfstatus, Hygiene) eine bedeutende Rolle im Hinblick auf den Gesundheitszustand zukommt. Im negativen Fall führte dieser Effekt dazu, dass die Anzahl auffälliger Befunde trotz vergleichsweiser guter Stallluftqualität (sehr) hoch war. Neben baulich-technischen Faktoren kommt folglich dem Management eine sehr große Bedeutung zu

Fragen des Arbeits-, Tier- und Umweltschutzes bei der Schweinemast in verschiedenen Systemen unter besonderer Berücksichtigung mikrobieller Belastungen

Vor dem Hintergrund häufiger Atemwegserkrankungen bei Schweinen und landwirtschaftlich Beschäftigten war es das Ziel eines interdisziplinären Verbundprojektes, Belastungen von Mensch, Tier und Umwelt in ökologischen und konventionellen Haltungssystemen für die Schweinemast zu erfassen. Die Untersuchungen wurden in zwei konventionellen (Stall A, B) (BVT-Stall, 50% reduzierter Schlitzanteil, Zwangsbelüftung) und zwei ökologischen Ställen durchgeführt: Stall C (Praxisstall: Tiefstreu, freie Fenster-Lüftung), Stall D (Praxisstall: EU-Öko-VO, Trauf-First-Lüftung). Über 2 Mastperioden (je 3 Messungen; kalte / warme Jahreszeit) wurden die Parameter erfasst: luftgetragene Endotoxine, Schimmelpilze, Bakterien mit Differenzierung, arbeitsmedizinische Staubfraktionen, Materialproben, Staubfraktion PM10, NH3, CO2 und CH4, Lufttemperatur, Luftfeuchte. Die Beschreibung der Tiergesundheit erfolgte über serologische (Mycoplasmen, PRRS-, Influenza-A- und Circo-Virus) und koprologische (Parasitenbefall) Analysen und über Schlachtkörper- und Organbefundungen (Lunge, Pleura, Perikard, Leber). Am Beispiel der Endotoxin-Konzentration wurden die z.T. sehr hohen biologischen Belastungen deutlich: Stall C (Median: 14.495 EU/m3), Stall A/B (5.544 EU/m3), Stall D (2.876 EU/m3). Personengetragene Messungen führten zu deutlich höheren Werten. Die CO2- und NH3-Konzentrationen lagen in allen Ställen im Durchschnitt deutlich unter dem Grenzwert von 3000 ppm bzw. 20 ppm. In Stall C und D war die CH4-Konzentration allerdings höher als in Stall A/B (oberer Bereich der Literaturangaben). Auffällige Lungenbefunde fanden sich bei ca. 45% der untersuchten Schlachttiere, unabhängig vom Haltungssystem; parasitäre Leberveränderungen wurden ausschließlich in den Ställen C und D ermittelt. Die Untersuchungsergebnisse unterstreichen die Bedeutung der Ausführung, Dimensionierung und Regelung des Lüftungssystems sowie die Sauberkeit bzw. Hygiene und insbesondere das Betriebsmanagement im Stall und ihre Schlüsselrolle hinsichtlich der Freisetzungsmengen von Schadstoffen sowie der Tiergesundheit. Demgegenüber kommt der Klassifizierung der Haltungsumwelt durch die Einteilung in Haltungssystemen nur eine geringe Aussagekraft zu

Multi-minicore Disease

Abstract Multi-minicore Disease (MmD) is a recessively inherited neuromuscular disorder characterized by multiple cores on muscle biopsy and clinical features of a congenital myopathy. Prevalence is unknown. Marked clinical variability corresponds to genetic heterogeneity: the most instantly recognizable classic phenotype characterized by spinal rigidity, early scoliosis and respiratory impairment is due to recessive mutations in the selenoprotein N (SEPN1) gene, whereas recessive mutations in the skeletal muscle ryanodine receptor (RYR1) gene have been associated with a wider range of clinical features comprising external ophthalmoplegia, distal weakness and wasting or predominant hip girdle involvement resembling central core disease (CCD). In the latter forms, there may also be a histopathologic continuum with CCD due to dominant RYR1 mutations, reflecting the common genetic background. Pathogenetic mechanisms of RYR1-related MmD are currently not well understood, but likely to involve altered excitability and/or changes in calcium homeoestasis; calcium-binding motifs within the selenoprotein N protein also suggest a possible role in calcium handling. The diagnosis of MmD is based on the presence of suggestive clinical features and multiple cores on muscle biopsy; muscle MRI may aid genetic testing as patterns of selective muscle involvement are distinct depending on the genetic background. Mutational analysis of the RYR1 or the SEPN1 gene may provide genetic confirmation of the diagnosis. Management is mainly supportive and has to address the risk of marked respiratory impairment in SEPN1-related MmD and the possibility of malignant hyperthermia susceptibility in RYR1-related forms. In the majority of patients, weakness is static or only slowly progressive, with the degree of respiratory impairment being the most important prognostic factor.</p

Altered splicing of the BIN1 muscle-specific exon in humans and dogs with highly progressive centronuclear myopathy

Amphiphysin 2, encoded by BIN1, is a key factor for membrane sensing and remodelling in different cell types. Homozygous BIN1 mutations in ubiquitously expressed exons are associated with autosomal recessive centronuclear myopathy (CNM), a mildly progressive muscle disorder typically showing abnormal nuclear centralization on biopsies. In addition, misregulation of BIN1 splicing partially accounts for the muscle defects in myotonic dystrophy (DM). However, the muscle-specific function of amphiphysin 2 and its pathogenicity in both muscle disorders are not well understood. In this study we identified and characterized the first mutation affecting the splicing of the muscle-specific BIN1 exon 11 in a consanguineous family with rapidly progressive and ultimately fatal centronuclear myopathy. In parallel, we discovered a mutation in the same BIN1 exon 11 acceptor splice site as the genetic cause of the canine Inherited Myopathy of Great Danes (IMGD). Analysis of RNA from patient muscle demonstrated complete skipping of exon 11 and BIN1 constructs without exon 11 were unable to promote membrane tubulation in differentiated myotubes. Comparative immunofluorescence and ultrastructural analyses of patient and canine biopsies revealed common structural defects, emphasizing the importance of amphiphysin 2 in membrane remodelling and maintenance of the skeletal muscle triad. Our data demonstrate that the alteration of the muscle-specific function of amphiphysin 2 is a common pathomechanism for centronuclear myopathy, myotonic dystrophy, and IMGD. The IMGD dog is the first faithful model for human BIN1-related CNM and represents a mammalian model available for preclinical trials of potential therapies

Cancer-testis antigen expression in triple-negative breast cancer

Background: Cancer-testis (CT) antigens, frequently expressed in human germline cells but not in somatic tissues, may become aberrantly reexpressed in different cancer types. The aim of this study was to investigate the expression of CT antigens in breast cancer. Patients and methods: A total of 100 selected invasive breast cancers, including 50 estrogen receptor (ER) positive/HER2 negative and 50 triple negative (TN), were examined for NY-ESO-1 and MAGE-A expression by immunohistochemistry. Results: A significantly higher expression of MAGE-A and NY-ESO-1 was detected in TN breast cancers compared with ER-positive tumors (P = 0.04). MAGE-A expression was detected in 13 (26%) TN cancers compared with 5 (10%) ER-positive tumors (P = 0.07). NY-ESO-1 expression was confirmed in nine (18%) TN tumor samples compared with two (4%) ER-positive tumors. Conclusions: MAGE-A and NY-ESO-1 CT antigens are expressed in a substantial proportion of TN breast cancers. Because of the limited therapeutic options for this group of patients, CT antigen-based vaccines might prove to be useful for patients with this phenotype of breast cance

Calcium Homeostasis in Myogenic Differentiation Factor 1 (MyoD)-Transformed, Virally-Transduced, Skin-Derived Equine Myotubes

Dysfunctional skeletal muscle calcium homeostasis plays a central role in the pathophysiology of several human and animal skeletal muscle disorders, in particular, genetic disorders associated with ryanodine receptor 1 (RYR1) mutations, such as malignant hyperthermia, central core disease, multiminicore disease and certain centronuclear myopathies. In addition, aberrant skeletal muscle calcium handling is believed to play a pivotal role in the highly prevalent disorder of Thoroughbred racehorses, known as Recurrent Exertional Rhabdomyolysis. Traditionally, such defects were studied in human and equine subjects by examining the contractile responses of biopsied muscle strips exposed to caffeine, a potent RYR1 agonist. However, this test is not widely available and, due to its invasive nature, is potentially less suitable for valuable animals in training or in the human paediatric setting. Furthermore, increasingly, RYR1 gene polymorphisms (of unknown pathogenicity and significance) are being identified through next generation sequencing projects. Consequently, we have investigated a less invasive test that can be used to study calcium homeostasis in cultured, skin-derived fibroblasts that are converted to the muscle lineage by viral transduction with a MyoD (myogenic differentiation 1) transgene. Similar models have been utilised to examine calcium homeostasis in human patient cells, however, to date, there has been no detailed assessment of the cells’ calcium homeostasis, and in particular, the responses to agonists and antagonists of RYR1. Here we describe experiments conducted to assess calcium handling of the cells and examine responses to treatment with dantrolene, a drug commonly used for prophylaxis of recurrent exertional rhabdomyolysis in horses and malignant hyperthermia in humans

Theranostic pretargeted radioimmunotherapy of colorectal cancer xenografts in mice using picomolar affinity 86Y- or 177Lu-DOTA-Bn binding scFv C825/GPA33 IgG bispecific immunoconjugates

Purpose: GPA33 is a colorectal cancer (CRC) antigen with unique retention properties after huA33-mediated tumor targeting. We tested a pretargeted radioimmunotherapy (PRIT) approach for CRC using a tetravalent bispecific antibody with dual specificity for GPA33 tumor antigen and DOTA-Bn–(radiolanthanide metal) complex. Methods: PRIT was optimized in vivo by titrating sequential intravenous doses of huA33-C825, the dextran-based clearing agent, and the C825 haptens [superscript 177]Lu-or [superscript 86]Y-DOTA-Bn in mice bearing the SW1222 subcutaneous (s.c.) CRC xenograft model. Results: Using optimized PRIT, therapeutic indices (TIs) for tumor radiation-absorbed dose of 73 (tumor/blood) and 12 (tumor/kidney) were achieved. Estimated absorbed doses (cGy/MBq) to tumor, blood, liver, spleen, and kidney for single-cycle PRIT were 65.8, 0.9 (TI 73), 6.3 (TI 10), 6.6 (TI 10), and 5.3 (TI 12), respectively. Two cycles of PRIT (66.6 or 111 MBq [superscript 177]Lu-DOTA-Bn) were safe and effective, with a complete response of established s.c. tumors (100 – 700 mm³) in nine of nine mice, with two mice alive without recurrence at >140 days. Tumor log kill in this model was estimated to be 2.1 – 3.0 based on time to 500-mm³ tumor recurrence. In addition, PRIT dosimetry/diagnosis was performed by PET imaging of the positron-emitting DOTA hapten [superscript 86]Y-DOTA-Bn. Conclusion: We have developed anti-GPA33 PRIT as a triplestep theranostic strategy for preclinical detection, dosimetry, and safe targeted radiotherapy of established human colorectal mouse xenografts.National Institute of Mental Health (U.S.) (Grant R01-CA-101830

CTdatabase: a knowledge-base of high-throughput and curated data on cancer-testis antigens

The potency of the immune response has still to be harnessed effectively to combat human cancers. However, the discovery of T-cell targets in melanomas and other tumors has raised the possibility that cancer vaccines can be used to induce a therapeutically effective immune response against cancer. The targets, cancer-testis (CT) antigens, are immunogenic proteins preferentially expressed in normal gametogenic tissues and different histological types of tumors. Therapeutic cancer vaccines directed against CT antigens are currently in late-stage clinical trials testing whether they can delay or prevent recurrence of lung cancer and melanoma following surgical removal of primary tumors. CT antigens constitute a large, but ill-defined, family of proteins that exhibit a remarkably restricted expression. Currently, there is a considerable amount of information about these proteins, but the data are scattered through the literature and in several bioinformatic databases. The database presented here, CTdatabase (http://www.cta.lncc.br), unifies this knowledge to facilitate both the mining of the existing deluge of data, and the identification of proteins alleged to be CT antigens, but that do not have their characteristic restricted expression pattern. CTdatabase is more than a repository of CT antigen data, since all the available information was carefully curated and annotated with most data being specifically processed for CT antigens and stored locally. Starting from a compilation of known CT antigens, CTdatabase provides basic information including gene names and aliases, RefSeq accession numbers, genomic location, known splicing variants, gene duplications and additional family members. Gene expression at the mRNA level in normal and tumor tissues has been collated from publicly available data obtained by several different technologies. Manually curated data related to mRNA and protein expression, and antigen-specific immune responses in cancer patients are also available, together with links to PubMed for relevant CT antigen article